Polarly localized EccE1is required for ESX-1 function and stabilization of ESX-1 membrane proteins in Mycobacterium tuberculosis

Soler-Arnedo, P.; Sala, C.; Zhang, M.; Cole, S. T.; Piton, J.

doi:10.1101/820324

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bioRxiv · 10.1101/820324

Polarly localized EccE1is required for ESX-1 function and stabilization of ESX-1 membrane proteins in Mycobacterium tuberculosis

Soler-Arnedo, P.·Sala, C.·Zhang, M.·Cole, S. T.·Piton, J.

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Abstract

Mycobacterium tuberculosis is a slow-growing intracellular bacterium with the ability to induce host cell death and persist indefinitely in the human body. This pathogen uses the specialized ESX-1 secretion system to secrete virulence factors and potent immunogenic effectors required for disease progression. ESX-1 is a multi-subunit apparatus with a membrane complex that is predicted to form a pore in the cytoplasmic membrane. In M. tuberculosis this complex is composed of five membrane proteins: EccB1, EccCa1, EccCb1, EccD1, EccE1. In this study, we have characterized the membrane component EccE1 and found that deletion of eccE1 lowers the levels of EccB1, EccCa1 and EccD1 thereby abolishing ESX-1 secretion and attenuating M. tuberculosis ex vivo. Surprisingly, secretion of EspB was not affected by loss of EccE1. Furthermore, EccE1 was found to be a membrane- and cell-wall associated protein that needs the presence of other ESX-1 components to assemble into a stable complex at the poles of M. tuberculosis. Overall, this investigation provides new insights into the role of EccE1 and its localization in M. tuberculosis.\n\nIMPORTANCETuberculosis (TB), the worlds leading cause of death of humans from an infectious disease, is caused by the intracellular bacterium Mycobacterium tuberculosis. The development of successful strategies to control TB requires better understanding of the complex interactions between the pathogen and human host. We investigated the contribution of EccE1, a membrane protein, to the function of the ESX-1 secretion system, the major virulence determinant of M. tuberculosis. By combining genetic analysis of selected mutants with eukaryotic cell biology and proteomics, we demonstrate that EccE1 is critical for ESX-1 function, secretion of effector proteins and pathogenesis. Our research improves knowledge of the molecular basis of M. tuberculosis virulence and enhances our understanding of pathogenesis.

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Soler-Arnedo, P., Sala, C., Zhang, M., Cole, S. T., Piton, J.. 2019-10-30. Polarly localized EccE1is required for ESX-1 function and stabilization of ESX-1 membrane proteins in Mycobacterium tuberculosis. https://doi.org/10.1101/820324

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